TRYPANOSOME REGULATORY PROTEIN

VSG基因在T.布氏杆菌含有多个表达位点 在阶段特异性细胞中高水平表达的相关基因 方式我们以前克隆和测序了一个ESAG，我们命名为T- LR（Trypanosome Leucine-rich Repeat）和其他人称之为ESAG 8。 从T-LR预测的蛋白质序列包含两个有趣的结构域。 一个与调节蛋白中的RING指基序具有同源性， 以序列特异性方式结合DNA，而另一种是富含亮氨酸的 在各种原核和真核生物中发现的重复序列（LRR） 参与蛋白质相互作用的蛋白质。因此， T-LR在寄生虫生理学中具有重要的调节作用。整体 本项目的目的是阐明T-LR的作用，并确定 其功能的分子机制。我们还计划确定和 表征T-LR可能相互作用的其他调节分子。 我们 将在细菌、酵母、昆虫或 哺乳动物表达系统，并产生针对 潜在的核酸和蛋白质结合域。 这些抗体将 用于表征T-LR蛋白的大小， 翻译修饰和阶段特异性表达，并确定 它的亚细胞位置。然后我们将识别出与 与锥虫细胞内的T-LR结合。这些核酸和蛋白质 他们的基因将被克隆。此外，我们将 调节T-LR基因的正常表达，通过敲除 T-LR基因在血液中的表达和T-LR基因在血液中的过度表达。 血流和前循环形式。 锥虫生长的变化， 形态学和其他特定基因的表达将鉴定细胞 涉及T-LR的过程。这些研究旨在确定新的 分子过程，为开发新的 寄生虫控制策略此外，与其他几项研究一样， 现象在T.他们还应该提供新的见解， 所有真核生物的基础分子生物学。

The VSG expression site of T. brucei contains several Expression Site Associated Genes which are expressed at high levels in a stage-specific manner. We have previously cloned and sequenced an ESAG which we named T- LR (for Trypanosome Leucine-rich Repeat) and others have called ESAG 8. The protein sequence predicted from T-LR contains two interesting domains. One has homology to the RING finger motif in regulatory proteins which bind DNA in a sequence-specific fashion, while the other is a leucine-rich repeat (LRR) sequence found in wide variety of prokaryotic and eukaryotic proteins involved in protein-protein interactions. Thus, it is likely that T-LR has an important regulatory role in parasite physiology. The overall aim of this project is to elucidate the role of T-LR and to identify the molecular mechanisms underlying its function. We also plan to identify and characterize other regulatory molecules with which T-LR may interact. We will express recombinant T-LR proteins in bacterial, yeast, insect or mammalian expression systems and produce specific antibodies against the potential nucleic acid- and protein-binding domains. The antibodies will be used to characterize the T-LR protein in terms of size, post- translational modification, and stage-specific expression and to determine its subcellular location. We will then identify molecules which interact with T-LR within the trypanosome cell. These nucleic acids and proteins will be characterized and their genes cloned. In addition, we will modulate the normal expression of the T-LR gene by knockout of the expressed T-LR gene in bloodstream forms and over-expression of T-LR in bloodstream and procyclic forms. Changes in trypanosome growth, morphology and expression of other specific genes will identify cellular processes involving T-LR. These studies are designed to identify new molecular processes that provide the basis for development of novel parasite control strategies. In addition, like the study of several other phenomena in T. brucei, they should also provide new insights into the fundamental molecular biology of all eukaryotic organisms.